Amorphous form of tetracyclic compound

ABSTRACT

An amorphous form of 9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile and a solid dispersion containing the amorphous form can be used extremely advantageously as drugs for oral administration.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of U.S. application Ser. No.16/425,603, filed May 29, 2019, which is a Divisional of U.S.application Ser. No. 15/501,267, filed Feb. 2, 2017, which issued asU.S. Pat. No. 10,344,014, on Jul. 9, 2019, which is the U.S. NationalStage application of PCT/JP2015/072450, filed Aug. 7, 2015, which claimspriority from Japanese application JP 2014-162899, filed Aug. 8, 2014.

TECHNICAL FIELD

The present invention relates to amorphous forms of tetracycliccompounds having inhibitory activity on ALK, and solid dispersionscontaining the amorphous forms.

BACKGROUND ART

Anaplastic lymphoma kinase (ALK) is one of receptor tyrosine kinasesbelonging to the insulin-receptor family (NPL 1 and NPL 2), and anabnormality in the ALK gene has been reported to lead to the productionof an abnormal kinase with the gene fused with another gene.

As diseases with abnormalities of ALK, cancer and cancer metastasis (NPL1 and PTL 1), depression, and cognitive function disorders (NPL 2) areknown. Accordingly, provision of ALK inhibitors will results in that ofeffective therapeutic and prophylactic agents against these diseases.

As compounds having inhibitory activity on ALK, a compound representedby the formula (I) (compound name:9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile)and the like are known (PTL 2, PTL 3, and PTL 4).

Since the compound is poorly soluble or insoluble to water, a sufficientbioavailability might not possibly be obtained when it is administeredorally.

CITATION LIST Patent Literature

-   PTL 1: Japanese Patent Laid-Open No. 2009100783(A)-   PTL 2: Japanese Patent No. 4588121-   PTL 3: Japanese Patent No. 4918630-   PTL 4: Japanese Patent Laid-open No. 2012-126711

Non Patent Literature

-   NPL 1: Nature, vol. 448, pp. 561-566, 2007-   NPL 2: Neuropsychopharmacology, vol. 33, pp. 685-700, 2008

SUMMARY OF INVENTION Technical Problem

With respect to the aforementioned problem, the present inventors found,as a result of extensive studies, that amorphous forms of hydrochloridesof the tetracyclic compound represented by the formula (I) below have avery high physical stability and an excellent solubility. Furthermore,the present inventors found that solid dispersions containing theamorphous forms have excellent physical and chemical stabilities andsolubility of the compound represented by the formula (I), a saltthereof, or a solvate of the compound or a salt thereof is improved.

Solution to Problem

More specifically, the present invention is as follows.

(1) An amorphous form of a compound represented by the formula (I):

a salt thereof, or a solvate of the compound or a salt thereof.(2) The amorphous form described in (1), wherein the amorphous formincludes a salt of the compound.(3) The amorphous form described in (1) or (2), wherein the salt is ahydrochloride.(4) The amorphous form described in any one of (1) to (3), wherein thesalt is a monohydrochloride.(5) The amorphous form described in any one of (1) to (4), wherein anexothermic peak is detected between 190°±5° C. and 230°±5° C. by adifferential scanning calorimetry analysis.(6) The amorphous form described in any one of (1) to (4), wherein aglass transition temperature is between 190±5° C. and 230±5° C.(7) The amorphous form described in any one of (1) to (6), wherein theamorphous form has an X-ray powder diffraction pattern shown in FIG. 1.(8) A method of producing an amorphous form of a compound represented bythe formula (I), a salt thereof, or a solvate of the compound or a saltthereof, the method including the steps of: (i) dissolving the compoundrepresented by the formula (I), the salt thereof, or the solvate of thecompound or the salt thereof into a solvent to prepare a feed solution;(ii) spraying the feed solution obtained in the step (i); and (iii)drying the sprayed feed solution to provide the amorphous form of thecompound represented by the formula (I), a salt thereof, or a solvate ofthe compound or a salt thereof.(9) The method described in (8), wherein the compound represented by theformula (I), the salt thereof, or the solvate of the compound or thesalt thereof in the step (i) is a crystalline form.(10) The method described in (8), wherein the compound represented bythe formula (I), the salt thereof, or the solvate of the compound or thesalt thereof comprises a hydrochloride of the compound represented bythe formula (I).(11) The method described in (9) or (10), wherein the compoundrepresented by the formula (I), the salt thereof, or the solvate of thecompound or the salt thereof comprises a monohydrochloride of thecompound represented by the formula (I).(12) The method described in (9) to (11), wherein the hydrochloride ofthe compound represented by the formula (I) in the step (i) is acrystalline form having an X-ray powder diffraction pattern with peaksat diffraction angles (2θ) of 8.4°±0.2°, 14.0°±0.2°, 16.7°±0.2°,18.8°±0.2°, and 23.3°±0.2°.(13) The method described in any one of (8) to (12), wherein the solventis tetrahydrofuran.(14) The method described in (13) wherein a concentration of thetetrahydrofuran is between 65% and 85%.(15) An amorphous form of a compound represented by the formula (I), asalt thereof, or a solvate of the compound or a salt thereof, whereinthe amorphous form is obtained by the method described in any one of (8)to (14).(16) An amorphous form of a monohydrochloride of the compoundrepresented by the formula (I), wherein the amorphous form is obtainedby the method described in any one of (8) to (14).(17) A composition including amorphous and crystalline forms of acompound represented by the formula (I), a salt thereof, or a solvate ofthe compound or a salt thereof.(18) The composition described in (17), wherein a content of thecrystalline form is 10% by weight or less.(19) The composition described in (17) or (18), wherein a content of thecrystalline form is 3% by weight or less.(20) The composition described in any one of (17) to (19), wherein acontent of the crystalline form is 1% by weight or less.(21) The composition described in any one of (17) to (20), wherein thecompound represented by the formula (I) or the salt thereof comprises ahydrochloride of the compound represented by the formula (I).(22) The composition described in any one of (17) to (21), wherein thecompound represented by the formula (I) or the salt thereof comprises amonohydrochloride of the compound represented by the formula (I).(23) The composition described in any one of (17) to (22), wherein theamorphous form is detected as an exothermic peak between 190°±5° C. and230°±5° C. by a differential scanning calorimetry analysis.(24) The composition described in any one of (17) to (22), wherein theamorphous form has a glass transition temperature of about 190±5° C. to230±5° C.(25) The composition described in any one of (17) to (24), wherein theamorphous form has an X-ray powder diffraction pattern shown in FIG. 1.(26) A solid dispersion including an amorphous form of a compoundrepresented by the formula (I), a salt thereof, or a solvate of thecompound or a salt thereof; and an inert carrier.(27) The solid dispersion described in (26), wherein the inert carrieris a solid polymer.(28) The solid dispersion described in (27), wherein the solid polymeris cellulose or a derivative thereof or a water-soluble syntheticpolymer.(29) The solid dispersion described in (28), wherein the solid polymeris hypromellose, hypromellose acetate succinate, hypromellose phthalate,carboxymethyl cellulose, carboxymethyl ethyl cellulose, sodiumcarboxymethyl cellulose, hydroxyethyl cellulose, hydroxyethyl methylcellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose,cellulose acetate phthalate, polyethylene glycol, polyvinyl alcohol,povidone, copolyvidone, polyvinyl acetate phthalate, polyvinylacetaldiethylamino acetate, cellulose acetate phthalate, a polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, anaminoalkyl methacrylate copolymer E, an aminoalkyl methacrylatecopolymer RS, a methacrylate copolymer L, a methacrylate copolymer LD, amethacrylate copolymer S, or a carboxyvinyl polymer.(30) The solid dispersion described in (29), wherein the solid polymeris selected from a polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol graft copolymer, hypromellose phthalate, hypromellose acetatesuccinate, and a methacrylate copolymer L.(31) The solid dispersion described in any one of (27) to (30), whereina weight ratio of the compound represented by the formula (I), the saltthereof, or the solvate of the compound or the salt thereof, as a freeform, and the inert carrier is 9:1 to 1:9.(32) A powder, fine granules, granules, a table, or a capsule containingthe solid dispersion described in any one of (26) to (31).

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a graph showing a result of X-ray powder diffractionmeasurement on an amorphous form of the present invention.

FIG. 2 is a graph showing a result of X-ray powder diffractionmeasurement on form I crystals.

FIG. 3 is a graph showing a result of X-ray powder diffractionmeasurement on form II crystals.

FIG. 4 is a graph showing a result of X-ray powder diffractionmeasurement on form III crystals.

FIG. 5 is a graph showing a result of differential scanning calorimetry(DSC) measurement on the amorphous form of the present invention.

FIG. 6 shows a polarizing micrograph of a compound A which has beendried after being completely dissolved in 70% THF (concentration of thecompound A: 14 mg/mL).

FIG. 7 shows a polarizing micrograph of the compound A which has beendried after being completely dissolved in 80% THF (concentration of thecompound A: 12 mg/mL).

FIG. 8 is a graph showing a result of X-ray powder diffractionmeasurement on the amorphous form and solid dispersions of the presentinvention.

FIG. 9 is a graph showing a result of X-ray powder diffractionmeasurement on amorphous forms and solid dispersions which have beenstored under conditions of 40° C. and 75% RH for 3 months.

FIG. 10 is a graph showing a result of X-ray powder diffractionmeasurement on amorphous forms and solid dispersions which have beenstored at 25° C. for 1 year.

FIG. 11 is a graph showing a result of a dissolution test on theamorphous forms and the solid dispersions of the present invention.

FIG. 12 is a graph comparing elution profiles of the amorphous forms andthe solid dispersions of the present invention with an elution profileof a crystalline pharmaceutical preparation of a hydrochloride.

DESCRIPTION OF EMBODIMENTS

An “amorphous form” as used in the present invention means a state ofsolid substances exhibiting no definite crystalline structure. Thepresence or absence of a crystalline structure can be examined bymeasuring X-ray powder diffraction (XRPD) under the measurementconditions given below. The amorphous forms in the present inventionhave a diffraction pattern containing a broad and weak peak (halo) whentheir X-ray powder diffraction (XRPD) was measured under the measurementconditions given below. The diffraction pattern becomes more distinctivewhen compared with those of crystalline forms. Glass-like substanceswhich have no crystalline structure and are similar to non-crystallinefluids with a very high viscosity are also included in the amorphousforms of the present invention.

Measurement conditions:

Measuring instrument: X'Pert-Pro MPD (manufactured by PANalytical)

Target: Cu

Tube voltage: 45 kV

Tube current: 40 mA

Step size: 0.02°

Scan axis: 2θ

Sampling time per step: 43 sec.

Scanning range: 3 to 40°

The presence of the amorphous forms of the present invention can beexamined using a well-known technique such as differential scanningcalorimetry (DSC), solid NMR, X-ray (powder) diffraction, IR, NIR, andRaman spectroscopy, optionally by comparing them with a crystalline formof a compound represented by the formula (I), a salt thereof, or asolvate of the compound or a salt thereof when they are mixed with thecrystalline form.

Specifically, for amorphous forms of monohydrochloride of the compoundrepresented by the formula (I) for example, an exothermic peak isdetected by a differential scanning calorimetry analysis between about190° C. and about 230° C. The term “about” means ±5° C., and preferably±2° C. In other words, the amorphous forms of the present invention havea glass transition temperature of about 190-230° C. The term “about”means ±5° C. Preferably, the glass transition temperature is about220-230° C.

When the X-ray powder diffraction is used, crystalline and amorphousforms can be discriminated easily from each other because the formerexhibits a diffraction pattern containing a sharp peak whereas thelatter exhibits a diffraction pattern containing a relatively broad andweak peak (“halo”). FIG. 1 shows an example of a result of X-ray powderdiffraction measurement on an amorphous form of a monohydrochloride ofthe compound represented by the formula (I).

When differential scanning calorimetry is performed, a differentialscanning calorimeter, a nitrogen gas as a purge gas, a rate oftemperature rise of 10° C./min, other devices and various DSCthermograms including conditions can be used. Crystalline and amorphousforms can be discriminated from each other because the crystalline formsare typically characterized by a sharp melt endothermic/endothermic peakand amorphous forms do not exhibit a specific endothermic peak that canbe found in the crystalline forms. Those skilled in the art can thusdetermine the presence of an amorphous form by comparing DSC thermogramsbetween the crystalline form and the amorphous form.

Some of the measurement conditions of the differential scanningcalorimetry are given below.

Instrument name: differential scanning calorimeter (DSC)

Model: Q200 (manufactured by TA Instruments)

Heating rate: 10° C./min. or 20° C./min.

Measurement temperature range: 25° C. to 350° C. (Provided that if thesample contains much residual solvent, a pre-treatment with heat at 25°C. to 125° C. or 25° C. to 175° C. may be required.)

Atmospheric gas: dry nitrogen

Flow rate of atmospheric gas: 50 mL/min.

Cell: aluminum pan (pinhole)

Sample amount: 5 mg to 10 mg

Reference standard: empty pan

The amorphous form of the present invention may be in a composition, inwhich the amorphous form is mixed with one or more crystalline forms.The amorphous form may be in a composition in which the amorphous formis mixed with the crystalline form at any mixing rate such as about 1%,2%, 3%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% by weight or lessrelative to the entire composition.

The content of the crystalline form contained in the composition can becalculated using, for example, an X-ray powder diffraction method.

On the other hand, the content of the amorphous form contained in thecomposition can be calculated using, for example, DSC.

Examples of crystalline forms of the compound represented by the formula(I) or salts thereof include crystalline forms (form I, II, and IIIcrystals) of monohydrochloride of the compound represented by theformula (I).

The form I crystals are characterized by having an X-ray powderdiffraction pattern with peaks at diffraction angles (2θ) of 8.4°,14.0°, 16.7°, 18.8°, and 23.3°. An example of a result of X-ray powderdiffraction measurement on the form I crystals is shown in FIG. 2, andexamples of peaks in X-ray powder diffraction patterns of these crystalsare shown in Table 1. The form I crystals can be obtained by adding asolution containing the compound of the formula (I) dropwise to a liquidmixture of ethanol and hydrochloric acid, which contains 1 molarequivalent or more of hydrochloric acid relative to the compound of theformula (I), while keeping the temperature of the liquid mixture atabout 35° C. or higher.

TABLE 1 Diffraction angle (2θ) 3.5 8.0 8.4 8.7 11.0 11.9 12.1 14.0 15.115.6 16.1 16.4 16.7 17.1 17.5 18.2 18.8 19.0 19.1 20.2 20.5 21.0 21.521.8 22.1 22.3 22.6 22.9 23.3 24.1 24.8 25.4 25.7 26.1 26.9 27.7 27.928.2 28.8 29.5 29.9 30.8 31.3 31.8 31.9 32.6 33.1 33.2 33.8 34.7 35.335.5 36.4 36.6 37.5 38.8 39.4

The form II crystals are characterized by having an X-ray powderdiffraction pattern with peaks at diffraction angles (2θ) of 9.2°,10.2°, 16.2°, 20.5°, and 21.6°, and more specifically at diffractionangles (2θ) of 9.2°, 10.2°, 16.2°, 17.5°, 19.5°, 20.5°, 21.6°, and22.8°. An example of a result of X-ray powder diffraction measurement onthe form II crystals is shown in FIG. 3, and examples of peaks in X-raypowder diffraction patterns of these crystals are shown in Table 2. Theform II crystals can be obtained by drying form III crystals describedlater at about 40° C. under reduced pressure.

TABLE 2 Diffraction angle (2θ) 3.6 6.6 8.9 9.2 10.2 11.1 11.3 12.0 12.113.3 14.6 16.0 16.2 17.3 17.5 18.0 18.2 18.5 18.8 19.5 20.0 20.5 20.821.6 22.2 22.8 23.2 23.8 24.2 24.6 24.8 25.6 26.2 26.9 27.5 27.8 28.428.9 29.2 29.6 30.3 31.0 31.3 32.6 32.9 33.6 34.0 34.8 35.5 36.2 37.738.3 38.8 39.4 39.9

In one embodiment of the present invention, the form II crystals aremonohydrate. The monohydrate herein is not specifically limited as longas it is a crystal that stably contains about 1 equivalent of waterunder environments (e.g., temperature and relative humidity) where drugsare generally stored and used.

The form III crystals can be obtained by adding a solution containingthe compound of the formula (I) dropwise to a liquid mixture of ethanoland hydrochloric acid, which contains 1 molar equivalent or more ofhydrochloric acid relative to the compound of the formula (I), whilekeeping the temperature of the liquid mixture at about 15° C. The formIII crystals are characterized by having an X-ray powder diffractionpattern with peaks at diffraction angles (2θ) of 12.7°, 14.3°, 15.0°,18.5°, and 25.7°, and more specifically at diffraction angles (2θ) of7.5°, 12.7°, 14.3°, 15.0°, 18.5°, 20.3°, 21.0°, and 25.7°. FIG. 4 showsan example of a result of X-ray powder diffraction measurement on theform III crystals, and Table 3 shows examples of peaks in X-ray powderdiffraction patterns of these crystals.

TABLE 3 Diffraction angle (2θ) 3.5 7.0 7.5 8.5 9.0 10.8 11.3 11.4 12.713.3 14.1 14.3 15.0 15.7 16.0 16.4 16.8 17.1 17.9 18.2 18.5 19.6 19.920.3 21.0 22.1 22.5 22.8 23.1 23.7 24.0 24.5 24.9 25.2 25.7 26.0 26.726.9 27.2 27.5 27.7 28.5 28.8 29.1 29.5 29.6 29.9 30.3 30.5 30.9 31.432.1 32.8 34.0 34.4 34.8 35.3 36.2 36.3 36.7 36.9 37.6 38.2

The analysis of crystalline forms using X-ray powder diffraction can beperformed according to a method usually used such as the “X-ray powderdiffraction method” described in, for example, Japanese Pharmacopoeia(16th edition). The same crystal form typically refers to crystallineforms of which diffraction angles 20 coincide with each other with anerror of ±0.2°.

An example of measurement conditions for an X-ray powder diffractionanalysis is given below.

Measuring instrument: X'Pert-Pro MPD (manufactured by PANalytical)

Target: Cu

Tube voltage: 45 kV

Tube current: 40 mA

Step size: 0.02

Scan axis: 2θ

Sampling time per step: 43 sec.

Scanning range: 3 to 40°

Water content of the crystals can be measured using a method usuallyused: for example, with a dynamic vapor sorption instrument or by theKarl Fischer method.

An example of measurement conditions for a dynamic vapor sorptioninstrument is given below.

Dynamic vapor sorption instrument: DVS-1 (Surface Measurement Systems)

Temperature: fixed temperature about 25° C.

Atmospheric gas: dried nitrogen

Flow rate of atmospheric gas: 200 sccm (mL/min.)

Minimum equilibration time: 10 min.

Maximum equilibration time: 1200 min.

An example of measurement conditions for the measurement of watercontent using a Karl Fischer analyzer is given below.

Method of analysis: coulometric titration method

KF analyzer: volumetric moisture meter, model KF-100 (manufactured byMitsubishi Chemical Corporation)

Anode solution: Aquamicron AX (manufactured by Mitsubishi ChemicalCorporation)

Cathode solution: Aquamicron CXU (manufactured by Mitsubishi ChemicalCorporation)

Examples of the salts of the compound represented by the formula (I) inthe present invention include hydrochlorides, hydrobromides,hydroiodides, phosphates, phosphonates, sulfates, and sulfonates such asmethanesulfonates and p-toluenesulfonates, carboxylates such asacetates, citrates, malates, tartrates, succinates, and salicylates, oralkali metal salts such as sodium salts and potassium salts; alkalineearth metal salts such as magnesium salts and calcium salts; andammonium salts such as ammonium salts, alkylammonium salts,dialkylammonium salts, trialkylammonium salts, and tetraalkylammoniumsalts. These salts are produced by contacting the compound with an acidor a base that can be used for the production of drugs.

A solvate of the compound represented by the formula (I) or a saltthereof can be either hydrate or non-hydrate. Examples of non-hydratesof the compound represented by the formula (I) or salts thereof includetheir solvates with alcohol (e.g., methanol, ethanol, and n-propanol),or dimethylformamide.

The amorphous forms of the present invention have an excellentsolubility and are thus expected to exhibit better absorbability inliving bodies, in particular, higher intestinal absorbability thanconventional crystalline forms. Accordingly, by using an amorphous formof the present invention in place of a crystalline form of the compoundrepresented by the formula (I) or a salt thereof, a daily dose can bereduced. It is useful as drugs and medicines, in particular, oralpharmaceutical formulations.

Although typical amorphous forms crystallize fast and may sometimes havea problem in stability, the amorphous forms of the present inventionhardly undergo crystallization and are thus stable. The amorphous formsof the present invention which are thus superior in stability areadvantageous in terms of manufacturing and providing as well as usingthe products with a certain level of quality as drugs.

The amorphous forms of the present invention can be manufactured usingspray drying.

The “spray drying” is a method of producing a dry powder of a compoundby atomizing a solution, slurry, or emulsion containing the compound,which is referred to as a “feed solution”, to fine droplets andinjecting them into a hot air stream. Injection of mist is also referredto as “spraying” and as a method for this, centrifugal spraying using arotary disk and pressure spraying using a pressure nozzle are wellknown.

Spray drying can be performed according to a method usually used. Withspray drying, it is possible to produce stable and uniform amorphousforms with a high reproducibility, which is advantageous in terms ofmanufacturing and providing products with a certain level of quality asdrugs.

Examples of the aforementioned spray drying include a production methodincluding the steps of: dissolving the compound represented by thegeneral formula (I):

a salt thereof or a solvate of the compound or a salt thereof into asolvent to prepare a feed solution; spraying the feed solution; anddrying the sprayed feed solution to give an amorphous form thereof.

One embodiment of the step of preparing the feed solution is to prepareit by dissolving form I crystals of monohydrochloride of the compoundrepresented by the formula (I) into a solvent.

Solvents used for the spray drying include tetrahydrofuran. Theconcentration of tetrahydrofuran is, for example, 65% to 85%, and about70% is preferable.

The temperature at which the compound represented by the formula (I), asalt thereof, or a solvate of the compound or a salt thereof isdissolved into a solvent is preferably about 50° C. (±5° C.).

The temperature during the spray drying is preferably about 120° C. (±5°C.).

If a sample obtained by spray drying contains a residual solvent, a stepof removing the solvent, such as a step of drying the sample underreduced pressure, can be included as a post-processing.

Furthermore, the present invention also relates to solid dispersionscontaining an amorphous form of the compound represented by the formula(I), a salt thereof, or a solvate of the compound or a salt thereof; andan inert carrier. Since the amorphous form of the compound representedby the formula (I), a salt thereof, or a solvate of the compound or asalt thereof is stabilized more in the solid dispersion, its storagestability is improved and thus it is useful for actual clinicalpractices.

The “solid dispersion” as used in the present invention refers to asolid composition in which one or more active ingredients are dispersedin an inert carrier or a matrix thereof. In the solid dispersion of thepresent invention, the active ingredient is preferably dispersed in theinert carrier or a matrix thereof at a molecular level.

The “inert carrier” as used in the present invention means a solid orliquid vehicle diluent and may contain a plurality of substances.Preferable examples of the inert carrier include solid polymers. Solidpolymers are not specifically limited as long as they arepharmaceutically acceptable and can keep an amorphous state of theactive ingredient(s). One of the solid polymers alone or a mixture oftwo or more can be used. A weight ratio of the amorphous form of thecompound represented by the formula (I), a salt thereof, or a solvate ofthe compound or a salt thereof, as a free form, to the inert carrier is,for example, 9:1 to 1:9, preferably 2:1 to 1:9, and more preferably 1:2to 1:9.

A preferable embodiment of the present invention includes a soliddispersion in which an amorphous form of the compound represented byformula (I), a salt thereof, or a solvate of the compound or a saltthereof is dispersed in a solid polymer. A more preferable embodimentincludes a solid dispersion in which the amorphous form is dispersed inthe solid polymer at a molecular level.

Examples of the solid polymer include celluloses and their derivatives,and water-soluble synthetic polymers.

Examples of celluloses and their derivatives include hypromellose,hypromellose acetate succinate, hypromellose phthalate, carboxymethylcellulose, carboxymethyl ethyl cellulose, sodium carboxymethylcellulose, hydroxyethyl cellulose, hydroxyethyl methyl cellulose,hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, andcellulose acetate phthalate. Among the aforementioned solid polymers,preferable examples include hypromellose acetate succinate andhypromellose phthalate.

Examples of water-soluble synthetic polymers include polyethyleneglycol, polyvinyl alcohol, povidone, copolyvidone, polyvinyl acetatephthalate, polyvinylacetal diethylamino acetate, cellulose acetatephthalate, a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolgraft copolymer (product name: Soluplus), an aminoalkyl methacrylatecopolymer E, an aminoalkyl methacrylate copolymer RS, a methacrylatecopolymer L, a methacrylate copolymer LD, a methacrylate copolymer S,and carboxyvinyl polymers. Among the aforementioned solid polymers,preferable examples include the polyvinyl caprolactam-polyvinylacetate-polyethylene glycol graft copolymer and the methacrylatecopolymer L.

These solid dispersions can be obtained preferably by spray drying.Specifically, it can be obtained by, for example, dissolving thecompound represented by the formula (I), a salt thereof, or a solvate ofthe compound or a salt thereof and other solid substance (such as thesolid polymer) in a solvent, spray drying the solution thus obtained atabout 100° C., and optionally, drying under reduced pressure as apost-processing.

The weight ratio of the solid polymer to the amorphous form of thecompound represented by the formula (I), a salt thereof, or a solvate ofthe compound or a salt thereof, as a free form, is, for example, 9:1 to1:9, preferably 2:1 to 1:9, and more preferably 1:2 to 1:9.

The amorphous forms of the present invention can be administered orallyor parenterally. As oral dosage forms, powders, fine granules, granules,tables, and capsules are suitably used. As parenteral dosage forms,suppositories and the like are suitably used. When the amorphous form ofthe present invention is used as a solid dispersion, the soliddispersion itself can be used as a pharmaceutical preparation. It canalso be used as a pharmaceutical preparation in the form of powders,fine granules, granules, tables, or capsules using a formulationtechnique such as compression molding. The amorphous forms, the soliddispersions and the pharmaceutical preparations of the present inventioncan be used for a liquid medicine in which they have been dispersed in apharmaceutically acceptable solution in advance. In such a case, theycan be used for syrups for oral administration or injections forparenteral administration which includes a lyophilized agent forinjection to be used after being reconstituted. They can also beprepared for liposomal drugs.

For preparation of the dosage forms, a coloring agent, a sweeteningagent, and a flavor, usually used; a diluent, an excipient, a binder, alubricant, a disintegrating agent, a softening agent, a suspendingagent, an emulsifying agent, a preservative, an antioxidant, asurfactant, a stabilizing agent, a pH adjusting agent, and a dispersingagent can be used. These various dosage forms may be prepared accordingto a method usually used, and may be prepared aseptically.

For example, the disintegrating agent can be starches such as cornstarches, potato starches, and partially pregelatinized starches,celluloses such as crystalline celluloses and microcrystallinecelluloses, low-substituted hydroxypropyl celluloses, croscarmellosesodium, crospovidone, carmellose calcium, carmellose sodium, andcarboxymethyl starch sodium.

As the excipients, examples include sugars such as lactose, sucrose,glucose, reducing maltose, mannitol, sorbitol, and erythritol, starchesand their derivatives such as corn starches, potato starches,pregelatinized starches, partially pregelatinized starches, dextrin, andpullulan, celluloses such as crystalline celluloses and microcrystallinecelluloses, magnesium silicate, magnesium aluminate metasilicate, sodiumhydrogen phosphate, calcium hydrogen phosphate, and talc.

Examples of the lubricants include carnauba waxes, hydrogenated oils,stearic acid, magnesium stearate, calcium stearate, talc, sodium stearylfumarate, and sucrose fatty acid ester.

Furthermore, the solid dispersion or the pharmaceutical preparation ofthe present invention may partly contain crystals of the compoundrepresented by the formula (I), a salt thereof, or a solvate of thecompound or a salt thereof. Preferably, at least 75% of the compoundrepresented by the formula (I), a salt thereof, or a solvate of thecompound or a salt thereof in the solid dispersion or the pharmaceuticalpreparation is present in the amorphous form, but not limited thereto.More preferably, this amount is at least 90%, and yet more preferably,95%. The percentage of the amorphous forms can be obtained by comparingspectra of the X-ray powder diffraction between the crystalline andamorphous forms.

Exemplified formulations of pharmaceutical preparations containing formI crystals of a monohydrochloride of the compound represented by theformula (I) as an active ingredient are shown in Table 4. Thesepharmaceutical preparations can be produced using the following method.

Ingredients to be formulated as granules are loaded and premixed in ahigh-shear granulator. An appropriate amount of purified water issprayed and the ingredients are agitated and granulated. They are thendried under vacuum to give dry granules. The dry granules are sizedusing a sizing machine. The sized granules thus obtained and ingredientsexternally added are mixed in a mixer to produce mixed powders. Themixed powders are filled in a capsule to produce a capsule drug.

The pharmaceutical preparation can contain 1%, 2%, 3%, 4%, 5%, 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, 96%, 97%, 98%, 99% by weight or less of the amorphous form ofthe present invention relative to the entire monohydrochloride of thecompound of the formula (I) contained in the pharmaceutical preparation.

TABLE 4 Prepa- Prepa- Name ration 1 ration 2 Ingredients to beHydrochloride of the compound of 161.33 161.33 formulated as formula (I)granules Lactose monohydrate 33.67 52.42 Hydroxypropyl cellulose 15.0015.00 Sodium lauryl sulfate 75.00 56.25 Carmellose calcium 15.00 0.00Low-substituted hydroxypropyl 0.00 15.00 cellulose IngredientsCarmellose calcium 28.35 0.00 externally Low-substituted hydroxypropyl0.00 28.35 added cellulose Magnesium stearate 1.65 1.65 Total 330.00330.00

In the present specification, the following abbreviations may be used.

MeOH: methanol

EtOH: ethanol

MeCN: acetonitrile

THF: tetrahydrofran

EtOAc: ethyl acetate

DMSO: dimethyl sulfoxide

tBuOH: tert-butyl alcohol

EXAMPLES

The present invention is described specifically with reference to theExamples but they are merely exemplified and the present invention isnot limited thereto.

Reference Example 1 Form I Crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloride

400 g of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilewas dissolved in a mixed solvent of 4.8 L of methyl ethyl ketone, 1.44 Lof acetic acid, and 1.68 L of distilled water at room temperature, andthe solution was added dropwise to a mixture of 12 L of ethanol and 0.8L of 2N hydrochloric acid at 60° C. Precipitated solids were recoveredby filtration, washed with 2 L of ethanol, and dried to give 357 g ofform I crystals of a monohydrochloride of the title compound.

Reference Example 2 Form III Crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloride

9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile(9.00 g) was dissolved in a mixed solvent of methyl ethyl ketone (90ml), distilled water (31.5 ml), and acetic acid (27.0 ml). This solutionwas added dropwise to a liquid mixture of ethanol (90 ml) and 2Nhydrochloric acid (18.00 ml) stirred at 15° C. while keeping thetemperature of the liquid mixture at 15° C. Subsequently, it was washedwith a mixed solvent of methyl ethyl ketone (18.00 ml), distilled water(6.30 ml), and acetic acid (5.40 ml) and then stirred at 15° C.Precipitated solids were recovered by filtration to give form IIIcrystals of a monohydrochloride of the title compound.

Reference Example 3 Form II Crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloride

The form III crystals obtained in Reference example 2 were washed withethanol (90 ml) and then dried under reduced pressure at 40° C. forabout 16 hours to give form II crystals of a monohydrochloride of thetitle compound.

Reference Example 4 Form II Crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloride

9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile(4.00 g) was added to a mixed solvent of methyl ethyl ketone (40 ml),distilled water (14 ml), and acetic acid (12 ml) and dissolved thereinat 35° C. This solution was added dropwise to a liquid mixture ofethanol (40 ml) and 2N hydrochloric acid (8.00 ml) (stirred at 15° C.)while keeping the temperature of the liquid mixture at 15° C. To thisliquid mixture, a mixed solvent of methyl ethyl ketone (8.00 ml),distilled water (2.80 ml), and acetic acid (2.40 ml) was added dropwisewhile keeping the temperature of the liquid mixture at 15° C.Subsequently, the liquid mixture was stirred at 15° C. Precipitatedsolids were recovered by filtration, washed with ethanol (40 ml) andthen dried under reduced pressure at 40° C. to give form II crystals(2.4805 g) of a monohydrochloride of the title compound.

Reference Example 5 Form III Crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloride

9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile(4.00 g) was added to a mixed solvent of methyl ethyl ketone (40 ml),distilled water (14 ml), and acetic acid (12 ml) and dissolved thereinat 35° C. This solution was added dropwise to a liquid mixture ofethanol (40 ml) and 2N hydrochloric acid (8.00 ml) (stirred at 15° C.)while keeping the temperature of the liquid mixture at 15° C. A mixedsolvent of methyl ethyl ketone (8.00 ml), distilled water (2.80 ml), andacetic acid (2.40 ml) was added dropwise while keeping the temperatureof the liquid mixture at 15° C. Subsequently, the liquid mixture wasstirred at 15° C. Precipitated solids were recovered by filtration togive form III crystals (7.8435 g) of monohydrochloride of the titlecompound.

X-Ray Powder Diffraction Analysis

X-ray powder diffraction was measured under the following conditions onform I, II, and III crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochlorideobtained in Reference examples 1 to 5. Results of the measurement on theform II, III, and I crystals are shown in FIGS. 2 to 4.

Measuring instrument: X'Pert-Pro MPD (manufactured by PANalytical)

Target: Cu

Tube voltage: 45 kV

Tube current: 40 mA

Step size: 0.02

Scan axis: 2θ

Sampling time per step: 43 sec.

Scanning range: 3 to 40°

[Example 1] Preparation of an Amorphous Form of Hydrochloride of theCompound of the Formula (I)

A mixed solvent (40 mL) of tetrahydrofuran (140 mL) and water (60 mL)was added to a vessel containing form I crystals (600 mg) ofmonohydrochloride of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileand the vessel was warmed in a water bath at 50° C. Subsequently, themixture was stirred until the salt was dissolved. The solution thusobtained was spray-dried at about 120° C. and dried under reducedpressure as a post-processing to give an amorphous form (530 mg) ofmonohydrochloride of the title compound.

Conditions during the spray-drying are as follows.

Spraying pressure: 20-30 mbar

Inlet temperature: 120° C.

[Test Example 1] X-Ray Powder Diffraction Analysis

X-ray powder diffraction was measured under the following conditions onthe amorphous form of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochlorideobtained in Example 1. A result of measurement is shown in FIG. 1.

Measuring instrument: X'Pert-Pro MPD (manufactured by PANalytical)

Target: Cu

Tube voltage: 45 kV

Tube current: 40 mA

Step size: 0.02°

Scan axis: 2θ

Sampling time per step: 43 sec.

Scanning range: 3 to 40°

The amorphous form obtained in Example 1 had an X-ray powder diffractionpattern with a halo pattern, confirming that it is an amorphous form.

[Test Example 2] Differential Scanning Calorimetry (DSC)

Differential scanning calorimetry was performed under the followingconditions on the amorphous form of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochlorideobtained in a manner similar to Example 1. A result is shown in FIG. 5.Two exothermic peaks (at 252.86° C. and 269.29° C.) were observed.Furthermore, after the exothermic change, the samples were removed atabout 300° C. and stored at room temperature. Then, X-ray powderdiffraction analysis was performed and as a result, it was found thattransformation from the amorphous form to the crystalline form hadoccurred. The glass transition temperature was found at about 224.5° C.

Name of instrument: differential scanning calorimeter (DSC)

Model: Q200 (manufactured by TA Instruments)

Heating rate: 10° C./min.

Measurement temperature range: 25° C. to 350° C.

Atmospheric gas: dried nitrogen

Flow rate of atmospheric gas: 50 mL/min.

Cell: aluminum pan (pinhole)

Sample volume: 5 mg to 10 mg

Reference standard: empty pan

[Example 2] Comparison of Solvents to Obtain Amorphous

Solubility of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloride(hereinafter, a compound A) to each solvent was evaluated.

5 mL of each of the solvents described in Table 5 was added to 5 mg ofform I crystals of the compound A. When undissolved crystals wereobserved, the mixture was stirred at room temperature and the crystalswere dissolved using ultrasound (step 1). After the processingdescribed, when undissolved crystals were observed, the mixture wasfurther stirred in a water bath at about 50° C. (step 2).

When the crystals were dissolved completely, the form I crystals of thecompound A were further added (step 3).

The procedure of the steps 1 to 3 was repeated.

For CH₂Cl₂, the step 2 and the later step were performed at roomtemperature.

Results obtained for the solvent other than THF are given in Table 6,and results obtained for THF are given in Table 7.

TABLE 5 Solvent used Water content (v/v) 0% 10% 20% 30% Solvent MeOH NANA NA X EtOH NA NA NA X Acetone NA NA NA X MeCN NA NA NA X THF X X X XEtOAC X NA NA NA CH₂Cl₂ X NA NA NA DMSO X tBuOH X Dioxane X NA: notassessed X: assessed

TABLE 6 Concentration of the compound A in solvent 1 mg/mL 2 mg/mL 4mg/mL 6 mg/mL 8 mg/mL 10 mg/mL Solvent 70% MeOH Y/— Y/— N/Y  — — — 70%EtOH Y/— Y/— Y/— N/Y  — — 70% acetone Y/— Y/— Y/— Y/— N/Y  — 70% MeCNY/— Y/— Y/— Y/— Y/— N/Y* EtOAc N/N  — — — — — CH2Cl2 N/— — — — — — DMSOY/— Y/— N/Y  — — — t-BuOH N/N  — — — — — 1,4-dioxane N/N  — — — — — Theresults obtained at room temperature are shown on the left side of “/”and the results obtained at about 50° C. while stirring are shown on theright side thereof. Results: Y: dissolved, N: not dissolved, —: nottested *Formation of solids was observed inside the test tube.

TABLE 7 Concentration of the compound A (mg/mL) 1 2 3 4 5 6 8 10 12 14Concentration of 100%  N/N  — — — — — — — — — THF 90% Y/— — Y/— — N/N —— — — — 80% Y/— — — Y/— — Y/— Y/— Y/— Y/— — 70% Y/— — — — — Y/— Y/— Y/—Y/— Y/— The results obtained at room temperature are shown on the leftside of “/” and the results obtained at about 50° C. while stirring areshown on the right side thereof. Results: Y: dissolved, N: notdissolved, —: not tested

[Example 3] Examination Using Polarizing Microscope

Among the solutions obtained in Example 2, those in which the form Icrystals of the compound A were found to be dissolved were dried onglass dishes placed on a hot plate heated at 100° C. or higher. Thesolids thus obtained were examined using a polarizing microscope.Results are shown in FIGS. 6 and 7. It was determined that amorphousforms could be obtained when 70% THF (in which the concentration of thecompound A is 14 mg/mL) and 80% THF (in which the concentration of thecompound A is 12 mg/mL) were used (see, FIGS. 6 and 7).

(Reference)

It was observed that when the form I crystals of monohydrochloride ofthe compound represented by the formula (I) were pulverized using a jetmill, a small amount of amorphous form was formed in crystals, but theamorphous form tended to crystallize soon.

[Example 4] Stability Test

The amorphous form of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloridewas stored under a certain condition and transition of the state of theamorphous form of the test compound was analyzed using X-ray powerdiffraction as in the Test example 1. Results are shown in Table 8. As aresult, it was found in a long-term stability test of 12 months underconditions of 30° C. and 75% RH and in an accelerated test of 6 monthsunder conditions of 40° C. and 75% RH that the amorphous form hardlycrystallized and physically stable, when it was stored in a sealedcondition.

TABLE 8 Storage temperature/humidity Storage period (month) (° C./% RH)Storage state 1 3 6 12 25/93 Open X X X — 30/75 Sealed — ◯ ◯ ◯ 40/75Sealed ◯ ◯ ◯ — ◯: no peak of diffraction angle, X: clear peak ofdiffraction angle, —: not measured

Example 5

A test of measuring intrinsic dissolution rate (according to USP chapter1087 “APPARENT INTRINSIC DISSOLUTION-DISSOLUTION TESTING PROCEDURES FORROTATING DISK AND STATIONARY DISK”) was performed on a compositioncontaining the amorphous and crystalline forms (form I crystals) of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochlorideat certain ratios. Results are shown in Table 9. It was concluded thatthe dissolution rate was increased when the content of the amorphous wasabout 15% or higher.

TABLE 9 Amount of amorphous (%) Intrinsic dissolution rate (mg/cm²/min)0 < Lower than detection limit 0.0078 5.6 0.0065 9.8 0.0065 16.8 0.0112

Example 6

Solubility of the amorphous form and form I crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloridewas examined using a shake flask method (37° C.). Results are shown inTable 10. It was found that the solubility of the amorphous form washigher than that of the form I crystals.

TABLE 10 Equilibration time Solubility (mg/mL) Sample solution (hour)Form I crystal Amorphous First solution for dissolution 1 0.0012 0.0045test (Japanese Pharmacopoeia, 24 0.0013 0.0025 16th edition) Simulatedintestinal fluid 1 <Lower than limit <Lower than limit (SIFsp) ofquantitation of quantitation 24 <Lower than limit <Lower than limit ofquantitation of quantitation Acetate buffer pH 4.5 1 0.0004 0.0005 24<Lower than limit 0.0004 of quantitation Phosphate buffer pH 8.0 1<Lower than limit <Lower than limit of quantitation of quantitation 24<Lower than limit <Lower than limit of quantitation of quantitationSolution for dissolution 1 0.3641 1.1506 test 24 0.4102 1.1552 Water 10.0286 0.0599 24 0.0251 0.0393 Solution for dissolution test: solutionobtained by adding 4% of surfactant (Triton-X100) to the first solutionfor dissolution test Limit of quantitation: 0.0004 mg/mL

Example 7 (Production of the Starting Material)

Crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilewere obtained using a method similar to the following method (which isdescribed in Example 805 of PTL 2).

500 g of6-cyano-2-(2-(4-ethyl-3-(4-morpholin-4-yl-piperidin-1-yl)phenyl)propan-2-yl)-1H-indole-3-carboxylicacid was dissolved in a mixture of 9.4 L of DMA, 270 mL of anhydrousacetic acid, and 1170 mL of DIPEA in a nitrogen gas stream and stirredat 90° C. for 1 hour. After being cooled at room temperature, 3.525 L ofmethanol and then 5.875 L of distilled water were added. Precipitatedsolids were collected through filtration, washed twice with 1.41 L of aliquid mixture of methanol:water=3:5 and then dried to give the titlecompound 389.6 g (85%).

(Preparation of Solid Dispersions of the Compound Represented by theFormula (I) by Spray Drying)

Solvents shown in Table 11 were added to vessels containing 2.58 g ofthe crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilethus obtained and 5.60 g of the solid polymer shown in Table 8 and themixtures were stirred until they were dissolved. The liquid mixturesthus obtained were spray dried at about 100° C. and dried under reducedpressure as a post-processing to give amorphous substances of thecompound represented by the formula (I) (Example 8) and soliddispersions (Examples 9 to 15).

Conditions during the spray drying are as follows.

Spraying pressure: 25-50 mbar

Inlet temperature: 100° C.

TABLE 11 Solid polymer and solvent used for dissolution Solvent Sol-Exam- compo- vent ple Solid polymer Manufacturer sition volume 8 None —THF 300 mL 9 Copovidone BASF THF 300 mL 10 Solplus BASF THF 300 mL 11Hypromellose Shin-Etsu THF 300 mL phthalate Chemical Co., Ltd. 12Hypromellose Shin-Etsu THF 300 mL acetate Chemical succinate Co., Ltd.13 Methacrylate Evonik Degussa THF:EtOH 320 mL copolymer L Japan (15:1)14 Povidone BASF THF:EtOH 320 mL (15:1) 15 Hypromellose Shin-EtsuTHF:H₂O 325 mL 2910 Chemical (12:1) Co., Ltd.

[Test Example 3] X-Ray Powder Diffraction Analysis

On the amorphous substances, the solid dispersions and the crystals asthe starting material, of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileobtained in Examples 8 to 15, X-ray powder diffraction was measuredunder the following conditions. Results of the measurement are shown inFIG. 8.

Measuring instrument: D8 discover with GADDS (manufactured by Bruker)

Target: Cu

Tube voltage: 40 kV

Tube current: 40 mA

Detection time: 100 sec.

Detection angle: phi 0°, chi 90°

Scanning range: 5 to 25.3°

Halo patterns were observed in all of the X-ray power diffractionpatterns of the solid dispersions obtained in Examples 9 to 15,indicating that they are amorphous forms.

[Test Example 4] Accelerated Test

Accelerated test was performed on the amorphous substances and the soliddispersions of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileobtained in Examples 8 to 15 under conditions of 40° C. and 75% RH for 3months, and X-ray powder diffraction was measured under the followingconditions. Results of the measurement are shown in FIG. 9. It wasconcluded that the solid dispersion hardly crystallized and physicallystable.

Measuring instrument: X'Pert-Pro MPD (manufactured by PANalytical)

Target: Cu

Tube voltage: 45 kV

Tube current: 40 mA

Step size: 0.02°

Scan axis: 2θ

Sampling time per step: 43 sec.

Scanning range: 5 to 35°

Test Example 5

The amorphous substances and the solid dispersions of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileobtained in Examples 8 to 15 were tested under two conditions: i) theywere stored in a thermostatic bath set at 40° C. for 3 months, and ii)they were stored in a thermostatic bath set at 40° C. and 75% RH for 3months in an open state. Residual rates of the principal drugs weremeasured under following conditions of high performance liquidchromatography. Results of the measurement are shown in Table 13. It wasconcluded that the amorphous substances and the solid dispersions werechemically stable.

Detector: UV absorptiometer (measurement wavelength: 230 nm)

Column: stainless column of 4.6 mm in inner diameter and 15 cm inlength, filled with 3.5 um of octadecylsilyl silica gel for liquidchromatography

Column temperature: fixed temperature about 35° C.

Mobile phase A: mixed solution of acetonitrile/trifluoroacetic acid(2000:1)

Mobile phase B: mixed solution of water/trifluoroacetic acid (2000:1)

Mobile phase supply: controlled concentration gradient with varyingmixing ratio of mobile phases A and B as follows.

TABLE 12 Time after loading Mobile phase Mobile phase (min.) A (vol %) B(vol %)  0-17 5 → 50 95 → 50 17-26 50 → 100 50 → 0   26-26.1 100 → 5   0 → 95 26.1-30  5 95 Flow rate: 1.0 mL/min. Loading amount: 10 μLSample temperature: fixed temperature about 25° C.

TABLE 13 Retention rate after 3 Retention rate after 3 months at 40° C.months at 40° C. in 75% RH Example 8 99.60 99.66 Example 9 99.66 99.21Example 10 99.79 99.71 Example 11 99.75 99.61 Example 12 99.71 99.60Example 13 99.63 99.59 Example 14 99.72 99.56 Example 15 99.84 99.73

[Test Example 6] Storage Stability Test

The amorphous substances and the solid dispersions of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileobtained in Examples 8 to 15 were subjected to storage stability testsat 25° C. for 1 year. X-ray powder diffraction was measured under thefollowing conditions. Results of the measurement are shown in FIG. 10.It was concluded that the solid dispersions hardly crystallized and werephysically stable.

Measuring instrument: X'Pert-Pro MPD (manufactured by PANalytical)

Target: Cu

Tube voltage: 45 kV

Tube current: 40 mA

Step size: 0.02°

Scan axis: 2θ

Sampling time per step: 43 sec.

Scanning range: 5 to 35°

[Test Example 7] Small-Scale Dissolution Test (R. Takano, et al., Pharm.Res. 23:1144-1156 (2006))

Solubility of the amorphous substances and the solid dispersions of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileobtained in Examples 8 to 15 in a fasted state simulated intestinalfluid (FaSSIF) was estimated using a small-scale dissolution tester(Vankel Technologies, Inc.) under conditions of 37° C. and a puddlerotation speed of 50 rpm. For each test sample, the concentration of thecompound of the formula (I) in the test solution was measured after 5,10, 15, 20, 25, 30, 45, 60, 120, and 240 minutes under the followingconditions of high performance liquid chromatography. Results of themeasurement are shown in FIG. 11.

Detector: UV absorptiometer (measurement wavelength: 337 nm)

Column: stainless column of 3.0 mm in inner diameter and 5 cm in length,filled with 3 um of octadecylsilyl silica gel for liquid chromatography

Column temperature: fixed temperature about 40° C.

Mobile phase A: mixed solution of water/trifluoroacetic acid (1000:1)

Mobile phase B: mixed solution of acetonitrile/trifluoroacetic acid(1000:1)

Mobile phase supply: controlled concentration gradient with varyingmixing ratio of mobile phases A and B as follows.

TABLE 14 Time after loading Mobile phase Mobile phase (min.) A (vol %) B(vol %)  0-1.5 65 35 1.5-1.51 65 → 5 35 → 95 1.51-3.0   5 95 3.0-3.01 5→ 65 95 → 35 3.01-3.5  65 35 Flow rate: 1.0 mL/min. Loading amount: 10μL

As a result, it was shown that the solubility of the amorphoussubstances of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrileis improved by formulating a polymer and that Soluplus (polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol graft copolymer)exhibited the largest effect and the hypromellose acetate succinate, thehypromellose phthalate, and the methacrylate copolymer L exhibited thesecond largest effect.

Example 16

All ingredients including the form I crystals of9-ethyl-6,6-dimethyl-8-(4-morpholin-4-yl-piperidin-1-yl)-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrilemonohydrochloridewere weighed at a formulation ratio shown in Table 15. These ingredientsexcept for magnesium stearate and some of sodium starch glycolate wereloaded in a high-shear granulator and mixed. Subsequently, wetgranulation was performed while adding purified water and the productswere subjected to wet sizing and dried under vacuum. The products werethen subjected to dry sizing to give granules. The granules thusobtained were mixed with magnesium stearate and the remainder of sodiumstarch glycolate in a V-shaped mixer to obtain granules for filling.

TABLE 15 Ingredients Example 16 (wt %) Hydrochloride of the compound offormula (I) 16.5% Lactose monohydrate 44.3% Crystalline cellulose 20.0%Sodium starch glycolate 6.0% Hydroxypropyl cellulose 5.0% Sodium laurylsulfate 7.7% Magnesium stearate 0.5%

Solubility of Example 16 in FaSSIF was estimated using a small-scaledissolution tester under conditions of 37° C. and a puddle rotationspeed of 50 rpm and compared with those of Examples 8 and 10. As aresult, as shown in FIG. 12, it was revealed that the solid dispersionusing the polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolgraft copolymer as a polymer carrier had an improved solubility ascompared with the crystalline pharmaceutical preparation of ahydrochloride.

[Test Example 8] Blood Level Measurement

Male beagle dogs purchased from Beijing Marshall Biotechnology Co., Ltd.were used in the test. The animals were not allowed access to food orwater 16.5 hours prior to the administration of the drug and injectedintravenously with famotidine 2 hours prior to the administration toadjust their stomach pH. The preparations obtained in Examples 10 and 16were each filled as a test compound in a gelatin capsule in an amount of5 mg/kg and administered orally. Blood samples were collected over timefrom 15 minutes to 24 hours after the administration of thepharmaceutical preparations from a cephalic vein in the forearm using asyringe treated with heparin, and were centrifuged to obtain plasma. Ablood level of the test compound was then examined (Table 16).

TABLE 16 Area under the curve Maximum blood concentration for bloodconcentration (ng/mL) (ng · hr/mL) Standard Standard Average deviationAverage deviation Example 10 567 205 2790 1030 Example 16 264 143 1140790

As compared with the pharmaceutical preparations in the crystallineform, the solid dispersions exhibited high blood levels of the testcompounds and it was found that intestinal absorbabilities were improvedby about 2.4 times based on the area under the curve for blood level.

As a result of the aforementioned investigation, the amorphous forms ofthe present invention were stable for a long time at normal temperatureand had an improved solubility. In addition, the solid dispersions ofthe present invention had excellent physical and chemical stabilitiesand further improved the solubility of the amorphous forms of thepresent invention. Accordingly, the amorphous forms and the soliddispersions of the present invention are extremely useful for drugs, inparticular, dosage forms such as oral drugs which often have problems inbioabsorbabilities.

1. A method of producing an amorphous form of a compound represented byformula (I):

a salt thereof, or a solvate of the compound or a salt thereof, themethod comprising the steps of: (i) dissolving the compound representedby the formula (I), the salt thereof, or the solvate of the compound orthe salt thereof into a solvent to prepare a feed solution; (ii)spraying the feed solution obtained in the step (i); and (iii) dryingthe sprayed feed solution to provide the amorphous form of the compoundrepresented by the formula (I), the salt thereof, or the solvate of thecompound or the salt thereof.
 2. The method according to claim 1,wherein the solvent is tetrahydrofuran.
 3. The method according to claim2, wherein a concentration of the tetrahydrofuran is between 65% and85%.
 4. An amorphous form of a compound represented by the formula (I),a salt thereof, or a solvate of the compound or a salt thereof, whereinthe amorphous form is obtained by the method according to claim 1.